Antibodies, recombinant antibodies, recombinant antibody fragments and fusions mediated plant disease resistance against fungi

Abstract

A method for the production of fungus resistant transgenic plants, plant cells or plant tissue comprising the introduction of an Ab, rAb, rAb fragment or fusion or vector of the invention or the vectors of the composition of the invention into the genome of a plant, plant cell or plant cell tissue and a transgenic plant cell comprising stably integrated into the genome a polynucleotide or vector of the invention or the vectors of the composition of the invention.

1. A polynucleotide comprising a nucleic acid sequence coding for a fusion protein, the fusion protein comprising
 a) an anti-fungal protein or peptide (AFP) and an antibody fragment, specifically recognising an epitope of an  Ascomyceta , and 
 b) a cellular targeting signal, 
 
       wherein the antibody fragment is a scFv encoded by Seq. ID. No. 1. 
     
     
       2. The polynucleotide of  claim 1 , further comprising a regulatory sequence for expression in a plant, plant organ, plant tissue, or plant cell. 
     
     
       3. A vector comprising the polynucleotide of  claim 1 . 
     
     
       4. An expression cassette comprising the polynucleotide of  claim 1 . 
     
     
       5. The polynucleotide according to  claim 2 , wherein the regulatory sequence is selected from the group consisting of constitutive, chimeric, tissue specific, inducible synthetic, and cryptic promoters. 
     
     
       6. The polynucleotide according to  claim 1 , wherein the AFP, and the antibody fragment are arranged in any order relative to each other. 
     
     
       7. A composition comprising a purified preparation of a polynucleotide comprising a nucleic acid sequence coding for a fusion protein, the fusion protein comprising
 a) an anti-fungal protein or peptide (AFP) and an antibody fragment, specifically recognising an epitope of an  Ascomyceta , and 
 b) a cellular targeting signal 
 wherein the antibody fragment is a scFv encoded by Seq. ID. No. 1. 
 
     
     
       8. A method for the production of Ascomyceta-resistant transgenic plants, plant cells, or plant tissues, the method comprising the introduction of a fusion protein into a genome of a plant, plant cell, or plant tissue, the fusion protein comprising
 a) an anti-fungal protein or peptide (AFP) and an antibody fragment, specifically recognising an epitope of an  Ascomyceta , and 
 b) a cellular targeting signal 
 wherein the antibody fragment is a scFv encoded by Seq. ID. No. 1. 
 
     
     
       9. A kit comprising a fusion protein, a polynucleotide coding for the fusion protein, or a combination thereof, with the fusion protein comprising
 a) an anti-fungal protein or peptide (AFP) and an antibody fragment, specifically recognising an epitope of an  Ascomyceta , and 
 b) a cellular targeting signal 
 wherein the antibody fragment is a scFv encoded by Seq. ID. No. 1. 
 
     
     
       10. The kit according to  claim 9 , wherein the antibody fragment comprises an enzyme for ELISA or immunoblot detection. 
     
     
       11. The kit according to  claim 10 , wherein the enzyme is an alkaline phosphatase or a horse radish peroxidase. 
     
     
       12. A polynucleotide comprising a nucleic acid sequence coding for a fusion protein, the fusion protein comprising
 a) an anti-fungal protein or peptide (AFP) and an antibody fragment, specifically recognising an epitope of an  Ascomyceta , and 
 b) a cellular targeting signal, 
 
       wherein the AFP is encoded by Seq. ID. No. 12. 
     
     
       13. The polynucleotide of  claim 12 , further comprising a regulatory sequence for expression in a plant, plant organ, plant tissue, or plant cell. 
     
     
       14. The polynucleotide according to  claim 13 , wherein the regulatory sequence is selected from the group consisting of constitutive, chimeric, tissue specific, inducible synthetic, and cryptic promoters. 
     
     
       15. A vector comprising the polynucleotide of  claim 12 . 
     
     
       16. An expression cassette comprising the polynucleotide of  claim 12 . 
     
     
       17. The polynucleotide of  claim 12 , wherein a scFv encoding sequence is linked to an AFP encoding sequence via Seq. ID. No. 19, and
 the scFv encoding sequence has the 5′ position with the AFP encoding portion in the 3′ position, or 
 the AFP encoding portion has the 5′ position and the and scFv encoding sequence has the 3′ position. 
 
     
     
       18. The polynucleotide according to  claim 12 , wherein the AFP, and the antibody fragment are arranged in any order relative to each other. 
     
     
       19. A composition comprising a purified preparation of a polynucleotide comprising a nucleic acid sequence coding for a fusion protein, the fusion protein comprising
 a) an anti-fungal protein or peptide (AFP) and an antibody fragment, specifically recognising an epitope of an  Ascomyceta , and 
 b) a cellular targeting signal 
 wherein the AFP is encoded by Seq. ID. No. 12. 
 
     
     
       20. A method for the production of  Ascomyceta -resistant transgenic plants, plant cells, or plant tissues, the method comprising the introduction of a fusion protein into a genome of a plant, plant cell, or plant tissue, the fusion protein comprising
 a) an anti-fungal protein or peptide (AFP) and an antibody fragment, specifically recognising an epitope of an  Ascomyceta , and 
 b) a cellular targeting signal 
 wherein the AFP is encoded by Seq. ID. No. 12. 
 
     
     
       21. A kit comprising a fusion protein, a polynucleotide coding for the fusion protein, or a combination thereof, with the fusion protein comprising
 a) an anti-fungal protein or peptide (AFP) and an antibody fragment, specifically recognising an epitope of an  Ascomyceta , and 
 b) a cellular targeting signal 
 
       wherein the AFP is encoded by Seq. ID. No. 12. 
     
     
       22. The kit according to  claim 21 , wherein the antibody fragment comprises an enzyme for ELISA or immunoblot detection. 
     
     
       23. The kit according to  claim 22 , wherein the enzyme is an alkaline phosphatase or a horse radish peroxidase.